Fig. 4
From: EGF receptor kinase suppresses ciliogenesis through activation of USP8 deubiquitinase
USP8 suppresses ciliogenesis by counteracting trichoplein degradation. a TetOn-RPE1 MBP-trichoplein-FLAG cells transfected with control or USP8 siRNA (#1 or #2) were cultured for 48 h in the presence or absence of doxycycline (Dox, 30 ng ml−1). b Sixteen hours after transfection with or without KCTD17 siRNA, RPE1 cells were further transfected with control or USP8 siRNA (#1 or #2), and then cultured for 32 h. c–e TetOn-RPE1 FLAG-USP8 WT (c, e) or S718A (d) cells were transfected with trichoplein (c, d; siTCHP, #1 or #2) or Aurora A siRNA (e; siAURKA). Six hours after transfection, these cells were treated with 100 ng ml−1 of Dox. Representative confocal images of acetylated-tubulin (green), FLAG (red) and DAPI (blue), percentages of ciliated cells, and immunoblotting analysis are shown. Normalized intensities of trichoplein/GAPDH are shown as mean from three independent biological replicates in b. Graphs represent mean ± SD from three independent biological replicates (n > 200 each). **p < 0.01, *0.01 < p < 0.05, n.s. not significant, two-tailed unpaired student’s t-tests. Scale bars, 10 μm
